AUBF As a remedy to be able to repeat the test:

● Detoxification of urine.

MIDTERMS ● Addition of acid in the form of hydrochloric

acid with concentration of 0.1m. ph is 6.0.
● Dialysis - urine is placed in a cellophane
PREGNANCY TEST bag and is immersed with clean running water.
Dialysis is separation of solid from liquid portion
using a semipermeable membrane. A method of
CLASSIFIED INTO FOUR (4) cleansing the urine (ie crystalloids are removed).
● Biological PT
○ Uses test animals RESULTS
○ The most accurate in producing results. Aside from the true pregnancy, we can obtain false
 Immunological PT positives.
 Chemical PT
 Clinician's Test ECTOPIC PREGNANCY
the fetus did not develop in the uterus, ie in the fallopian tube
('dating place' of sperm and egg). After fertilization in the
The main purpose of PT is to determine the normal fallopian tube, the fetus is stuck.
pregnancy. ● Physicians suggest abortion. if carried for 9 months.
➔ The woman becomes pregnant with the development ● Sometimes the baby survives, sometimes both die.
of the embryo coming from the union of egg and
sperm. Later on, it becomes a fetus and when
delivered, it is called a baby. CAUSE OF FALSE POSITIVES:
➔ Period of 9 months ● When the female patient has increased
➔ PT is also done to differentiate true pregnancy from FSH (follicle stimulating hormone).
other conditions such as hydatidiform mole (latu latu - ● When a female patient has appendicitis.
agar).
➔ Even if it is not a fetus, the feeling of the female would
● Has incomplete abortion - there are certain traces of
tissues left in the uterus (endometrial lining). Can
be as if they are pregnant
result in bleeding. Do the DNC. Dilatation and
(cravings, nausea and vomiting).
curettage.
➔ In the case of male patients, it is to determine whether
they have testicular embryonal carcinoma (teratoma).
-seasonal variation of test animals
Also tumor of the pituitary gland.
-if the test animal is pregnant, the natural result is
positive.
The principle depends upon the hormone, hCG. can be
found in blood in highest concentration. Use specimens
with 2nd highest concentration as blood extraction is
BIOLOGICAL PREGNANCY TEST
refused. It is urine.
Aschheim-zondek
THE HORMONE CAN ALSO BE FOUND IN ● Use of white mice
● (CSF) Cerebrospinal fluid ● Immature female white mice. age is estimated.
● Saliva should be 21-30 days old.
● Sweat ● Quantitative test
● Tears ● Purpose is to differentiate true pregnancy from
● chorioepithelioma.
Measurement of the hormone is in international units or ● A 24 hour urine sample is being injected into the
mouse unit. Gonadotropin activity of 0.1mg of hCG subcutaneous of the white mice. After about 48
preparation required to produce cornification of the vaginal hours, sacrifice the mice and do an autopsy. Dissect
epithelium of an immature female rat. the animal and observe the ovary of the test animal.
Observe corpora hemorrhagica (red spots), Corpora
 2 lines = positive lutea (yellow spots). This indicates a positive result.
 1 line = negative
Friedmann’s Test
If positive with males, (false positive) patients may have ● Quantitative test
abnormal conditions, for females hydatidiform mole. ● Used for suspected hydatidiform, or teratoma.
● Principle is when hCG is injected to a
● Patient should avoid drugs with hCG female virgin rabbit, it will produce 1-6 or more
● or with toxic effects or tested with test corpora hemorrhagic on the surface of the ovary.
animals. When the ovary becomes full, we can observe
corpora lutea.
If urine is alkaline, it might kill the test animal.
In the lab, female and male cages are separated. Animals
Death of test animal may be due to: are born there and as they are separated, one will know if
● Toxic Substances animals are virgins or not. As for the rabbit, observe the
● Alkaline Urine behind. If shaped like a v, it is a virgin. if shaped like a u, it
● Bacterial Contamination
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 ● Prostigmin is injected to the patient, if a woman is
is used.
going to menstruate 2 weeks after the procedure, she
is not pregnant.

Hoffmann’s Test
● Also makes use of female virgin rabbit Skin Colostrum Test
● Specimen is serum ● First secretion of milk by the mammary glands.
● Injected into the marginal ear of the rabbit
● Colostrum is injected subcutaneously in the forearm
● Same result observation in Friedmann's of the patient.
Test.
● Similar procedure to antibiotics. Small amount of it
should be injected to observe for allergic reaction.
Frog Test
Two (2) Types ● If the area swells and shows reddish coloration, not
pregnant.
1. Hogbem's Test ● If there is no swelling and reddish coloration, she is
● using female frog pregnant.

Principle: ➔ Colostrum is secreted only by females who are pregnant

when hCG is injected into a female frog, it will cause
excretion of eggs or ova. The urine is injected into the thigh
muscle of the female frog. Set aside the frog for about 8-12
hours. During this period, do not feed the frog. 12 hours after
the urine of the frog is collected, using a syringe without a
needle, position in the urethra of the frog and extract. Urine
is placed on the glass slide. Look for eggs or ova of the frog.
2. Gallimainini Test
● using male
Principle:
When hCG is injected to a male frog, it causes the release of
cigar shaped spermatozoa. Same procedure as above.
If there is no hCG in the patient sample, there is no egg or
sperm produced by frogs
 Xenopus laevis (South african frog) - female frog
 Rana vittigera and Rana pipiens (green grass
frog) - male frogs
CHEMICAL PREGNANCY TEST
➔ Less accurate, less popular
➔ Mixture of certain reagents to the urine sample.
Pregnanediol Test
 Detects progestene.
 Color is observed, deep
orange to dark brown is
positive.
 Few amounts are added.
Iodine or Lugol’s Reaction Test
● Detects the presence of epinephrine.
● Positive is deep orange to deep brown.
Cytotrophoblast cell of the placenta produces hCG hormone.
Found in highest concentration in serum, 2nd in urine and
next in CSF.
CHEMICAL PREGNANCY TEST
Cannot be done outside the physician’s clinic
Prostigmin Withdrawal Bleeding Test ??
or delivered. If not, colostrum is a foreign substance to the
body and allergic reactions will manifest.
Primodos oral
● Comes in the forms of tablet
● Physician gives two tablets
● Taken once a day for two consecutive days
● Total of four tablets
● Observe five days after. If a woman menstruates,
she is not pregnant. If she does not menstruate, she is
pregnant.
Gestex
● These tablets are taken in the same manner as
primodos oral.
● Only one tablet is taken for a day only.
● After 45 days, if the patient menstruates, she is not
pregnant.
Back in the olden days, there were many women who had
wrong interpretations about these procedures. They thought
that gestex would abort the baby.
IMMUNOLOGICAL PREGNANCY TEST
Makes use of antigen or antisera, same procedure in blood
typing.
➔ Kit: vial of antisera, dropper (sometimes there is filter
inside), slide (sometimes made of glass or glossy
material similar to glossy camera film).
➔ Drop of antisera is dropped next to the drop of urine,
then mixed by swirling the slide.
➔ If agglutination occurs, positive for direct AT
➔ Indirect AT: if there is agglutination, negative. If not,
positive.
➔ Specimens should be freshly voided. Not refrigerated,
not preserved.
➔ Antigen/antisera should be refrigerated. Before using
vial should be rubbed with palms
➔ IPT is not 100% accurate
➔ The IPT result will show in about 2 minutes.
➢ Biological is most popular as it is accurate.

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In the Mid and late 90s, the test kit was hexagonal. There is
a specific area for drop of urine. There will be a plus sign
observed for positive and minus sign for negative.
A patient who would undergo a pregnancy
test should avoid drugs with hCG or other substances that
may affect the result.
(CSF) CEREBROSPINAL FLUID
 Not collected by medtech; only those who are
trained
 Clear and colorless as distilled water
➔ NEEDLE used is not same as the needle in blood
collection (Needle is much longer)
➔ COLLECTION is done in between the 3rd or 4th
Lumbar Bone (between the SPACE, not on the
Bones or Veins).
 Choroid Plexus – between the 3rd and 4th lumbar
space.
pH of CSF
 Normally slightly Alkaline (7.30 – 7.45)
Specific gravity
 1.006 – 1.008
Specific gravity
 Normally Clear and Colorless, akin to distilled
water.
 If there is a Bright Red Color (Fresh Blood) – it
might due to puncturing of a vessel (vein) during the
insertion of the needle.
 If there is a Brownish Red Color (Old blood) –
there might be intracranial hemorrhage.
 If the color is Yellow Xanthochromic – it might be
due blood pigments (RBC disintegration).
 If the color is Greenish/ Grayish – it is due to pus
cells; severe inflammatory reaction; acute
meningitis.
Note:
 Transparency of CSF = Clear
 Number of WBC < 200 /mm3
Coagulation
 CSF does not usually coagulate.
 But whenever coagulation is observed, it is
“clinical significant”.
 If the coagulation is described as “Kabuet” (pine
tree or web like); it indicates Tuberculosis
meningitis.
 If there is heavy coagulum + sediments occur; it is
seen in Acute suppurative meningitis.
 Complete coagulation with Xanthochromia but
without hemorrhage; occurs in Froin’s syndrome
(refers to spinal sub-arachnoid block).
 Test to confirm the presence of Sub-arachnoid
block is known as Queckenstedt Test.
Prodcution of CSF
 CSF is produced in the Choroid Plexuses of the
Lumbar ventricles and the 3rd and 4th ventricles of
Lumbar region.
 Usually, 20 ml of CSF is produced by the Adult
every hour.
 Adult produces 20 – 150 ml of CSF.
 Infants produces 10 – 60 ml of CSF.
NOTE
➔ Backbone is joined together by Fibro (fibrous)
cartilage. Whereas, the backbone has 5 lumbar
bones.
➔ Skull has a layer of CSF that protects the Brains.
However, in collection of CSF, skull is the Last
option.
 CSF specimen should be treated and
handled carefully as;
1. It might contain organisms that can
cause disease (infectious).
2. It is not easily collected.
➔ In the Brain there are 3 layers.
I. Dura mater – Outer layer, lining the skull and
the vertebral canal.
II. Arachnoid mater – inner membrane.
III. Pia mater – thin membrane that lies the
surfaces of the brain and spinal cord.
Collection of CSF
 CSF specimen is always a priority (STAT), as it
does not contain enough Fibrinogen to form a clot.
 However, if the CSF is not in a STAT basis such as
for the following;
a. Hematologic exam - the tube has to
be Refrigerated.
b. Microbiology – tube has to remain in
a room temperature.
c. Chemistry and Serology – tube has
to be frozen.
 After collecting the CSF in the syringe, it will be
placed into 3 labelled tubes.
Tube 1 (Base): Use for Chemical and
Serological Tests.
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 Tube 2 (Middle): Use for Microbiological  Sodium Fluoride - prevent glycolysis by inhibiting
examination (Free of microorganisms). the enzymatic activity (1 mg of fluoride per mL of
Tube 3: Use for Cell count – Wherein, CSF).
immediate processing is required since WBCs  1 mg NaF / 1 ml of CSF (1:1)
and RBCs begins to lyze within 1 hour. But, if Macroscopic Examination
the specimen cannot be analysed, it has to be - Measure the volume of the specimen (especially
Refrigerated. Since, if the specimen is left body fluids)
standing for about 2 hrs, about 40% of WBCs - Normal Volume of CSF = 1cc / lb (pound) of body
will disintegrate. weight
Xanthochromia - Daily production of CSF volume is roughly from
- Describing the color of the specimen as yellow, 100 – 150 cc.
pink, or orange.
NOTE

Factors that cause Xanthochromia  Increase amount of the CSF volume is observed in
1. Present of RBC degradation products. acute and chronic congestion of the meninges
2. If the color is PINK, it has a very slight amount of (membrane).
Oxyhemoglobin.  Increase amount in acute and chronic infections,
3. If the color is ORANGE, it has a heavy Hemolysis. especially when there is production of Exudates
4. If the color is YELLOW, it indicates the conversion (liquids with inflammation) and Transudates
of Oxyhemoglobin to Unconjugated bilirubin. (liquids without inflammation process).
5. Elevated serum bilirubin, carotin, increase
protein concentration, and melanoma pigment. Chemical Examination for CSF
➔ Qualitative Tests for Proteins (specifically for
NOTE GLOBULINS):
 The present of increase bilirubin signifies 1. Nonne - Apelt Test (Globulins)
IMMATURE LIVER FUNCTION. 2. Ross - Jones Test (Globulins)
 When the CSF is grossly bloody, Intracranial 3. Pandy’s Test (Albumin and Globulins)
hemorrhage is observed. 4. Noguchi’s Test (Protein)
 If there is really a hemorrhage, there is a way to 5. Benedict’s Test (Glucose/Sugar)
determine whether it is resulting from a Quantitative Tests:
hemorrhage or just from a traumatic tap (insertion 1. Nelson – Somogyi Test (Glucose / Sugar)
of the needle to the lumbar space), there are 3 - Use in Clinical Chemistry
Visual examinations. 2. Schales and Schales Test
1. Distribution of Blood = - Test for Chlorides
a. Uneven distribution = Traumatic Tap 3. Lange’s Colloidal Gold Test
b. Even distribution = Cerebral hemorrhage - Albumin / Globulin Ratio Test
2. Clot Formation - Uses 10 test tubes to make dilutions.
a. With Clot Formation = Traumatic Tap Procedure/s:
b. NO Clot Formation = Cerebral o 10 test tubes are needed + 2 tubes (as
(intracranial) hemorrhage control).
3. Xanthochromic Supernatant o on the 1st tube, 0.9 cc of 0.4% sodium
- Hemolysis becomes noticeable when RBC remain chloride solution is placed.
in the CSF for at least 2 hrs. o Then, 0.5 cc of the solution is transferred
- For the recent hemorrhage, will produce a clear into the other tubes up to the 11th tube.
supernatant and when there is introduction of o Place 0.85 cc of 1% NaCl in the 12th tube.
serum protein it may cause xanthochromic
o Add 0.1 cc of CSF in the 1st tube then
specimen.
mixed it well by sucking the fluid into the
Preservation of CSF
pipette (4x).

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 o After mixing, 0.5 cc will be transferred to
the 2nd tube (repeat in each successive
tube including the 10th tube).
o But the 11th and 12th tube are not included.
o to each of the 12 tubes, add 2.5 cc of
colloidal gold solution.
 Leutic curve shows the marked change in 3 rd and
4th tube.
 Meningitis curve shows the marked change in the
last 3 tubes.

NOTE
 11th tube serves as a control for the Stability of
the colloidal gold.
 12th tube serves as a control for the Sensitivity of
the colloidal gold. It should become colorless
within an hour.

 Let it stand overnight and read the result the next PREGNANCY TEST
morning. In which, color changes should be
observed. .
 Reporting should be in order (sequential)
Interpretation:
Unchanged Deep Red – 0
Red to Blue – 1
Lilac to Purple – 2
Deep Blue – 3
Pale Blue – 4
Colorless – 5

NOTE
 Normal Reaction – all tubes (1-10) is
unchanged.
 Paretic Curve (Zone 1) – 1st 3 tubes has
Increase number (ex. 5, 5, 5, 5, 5, 4, 3, 0, 0, 0).
It is seen in insane and multiple myeloma
patients.
 Leutic / Tabetic Curve (Zone 2) – (ex. 0, 0, 0,
3, 3, 3, 3, 0, 0, 0). It is seen in syphilitic
patients.
 Meningitis (Zone 3) – (ex. 0, 0, 0, 0, 2, 4, 4, 4,
4, 0). It is seen in Tuberculosis meningitic
patients.
 Curve in Brain tumor and Tuberculosis
meningitis patient – (ex. 0, 0, 0, 0, 2, 4, 3, 3,
3, 3).

NOTE

 Paretic curve shows the marked change in the 1 st

3 tubes.

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